Enhanced Expansion of Tumor-Infiltrating Lymphocytes for Adoptive Cell Therapy

Abstract

Disclosed herein is a method for ex vivo expanding tumor-infiltrating lymphocytes for use in adoptive cell therapy (ACT). The method involves culturing tumor fragments from the subject in a culture medium containing IL-2 and a 41BB agonist in an amount effective to expand tumor-infiltrating lymphocytes with enriched tumor-reactivity and specificity. Also disclosed is a method for treating a tumor in a subject that involves treating the subject with nonmyeloablative lymphodepleting chemotherapy, and administering tumor-infiltrating lymphocytes expanded by the disclosed methods.

1 .- 29 . (canceled) 
     
     
         30 . A method for improving ex vivo rapid expansion (REP) of tumor-infiltrating lymphocytes (TILs), the method comprising culturing the TILs in a two-stage process:
 (a) culturing tumor fragments from a human subject in a pre-REP first cell culture medium comprising IL-2 and a 4-1BB agonist antibody;   (b) removing at least a plurality of the TILs; and   (c) expanding the TILs in a REP second cell culture medium.   
     
     
         31 . The method of claim  1 , wherein the 4-1BB agonist antibody is present in the first medium at a concentration in the range of 0.5 μg/mL to 10 μg/mL. 
     
     
         32 . The method of claim  1 , wherein the 4-1BB agonist antibody is further added to the second culture medium at least every three days during step (c). 
     
     
         33 . The method of claim  1 , wherein the first culture medium further comprises a checkpoint inhibitor selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or a combination thereof. 
     
     
         34 . The method of claim  1  further comprising selecting a subpopulation of the ex vivo expanded TILs that are CD8 +  and PD-1 +  from the total population of ex vivo expanded TILs. 
     
     
         35 . The method of claim  1 , wherein the culturing in the pre-REP stage is for about 1 week to about 3 weeks. 
     
     
         36 . A method for treating a tumor in a subject, comprising the steps of:
 (a) expanding tumor-infiltrating lymphocytes from a tumor fragment from the subject using the method of claim  1 ;   (b) treating the subject with nonmyleoablative lymphodepleting chemotherapy; and   (c) administering the tumor-infiltrating lymphocytes to the subject.   
     
     
         37 . The method of claim  7 , wherein the IL2 is present in the first medium at a concentration of 3000 IU/ml or 6000 IU/mL. 
     
     
         38 . The method of claim  7 , wherein the subject is treated with anti-PD1 prior to the surgical resection of the tumor fragment. 
     
     
         39 . The method of claim  7 , further comprising, prior to the culturing, enzymatically digesting the tumor fragments using a digest mixture. 
     
     
         40 . The method of claim  10 , wherein the digest mixture comprises a deoxyribonuclease (DNase), a collagenase, a hyaluronidase, or a combination thereof. 
     
     
         41 . The method of claim  7 , wherein the expanded TILs comprise T cells with a phenotype selected from the group consisting of CD3 + CD4 + , CD3 + CD8 + , CD8 + CD28 + , CD8 + CD27 + , CD8 + CD27 + CD28 + , and combinations thereof. 
     
     
         42 . The method of claim  7 , wherein the subject is a human.